Use of soluble African horse sickness viral protein 7 as an antigen delivery and presentation system
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Date
Authors
Rutkowska, Daria Anna
Meyer, Quinton Christian
Maree, Francois Frederick
Vosloo, Wilna
Fick, Wilma
Huismans, H. (Henk), 1942-
Journal Title
Journal ISSN
Volume Title
Publisher
Elsevier B.V.
Abstract
We have investigated the use of soluble chimeric trimers of the major capsid protein VP7 of
African horse sickness virus (AHSV) as a vaccine delivery system by targeting some of the
natural hydrophilic loops on the VP7 top domain for the insertion of foreign peptides. Key to this
trimer display strategy is the solubility of AHSV VP7 and how the solubility of this hydrophobic
protein can be manipulated by inserting peptides into the top domain. To investigate, we
generated different cloning vectors by inserting multiple cloning sites at three different positions
in the VP7 gene. These modifications inserted six amino acids at the cloning sites and in some
cases this converted VP7 to a largely soluble protein without affecting the ability of the modified
proteins to form trimers. The vectors were used to generate a number of soluble VP7 fusion
proteins including a fusion with a 36 amino acid insert that overlaps important immunological
domains on protein VP1 of foot and mouth disease virus (FMDV) as well as a 110 amino acid
peptide derived from AHSV VP2. Soluble trimers of these fusion proteins were able to elicit a
good insert-specific immune response in guinea pigs. l-Arginine was found to reverse protein
aggregation and was employed as an effective strategy to isolate relatively pure soluble chimeric
VP7 trimers. Another factor that increased VP7 solubility in both wild-type VP7 and one of the
VP7 vector proteins was the substitution of the leucine residue in position 345 of the VP7 Cterminus
with a hydrophilic arginine residue.
Description
Keywords
Immune display, Soluble VP7 trimers, Fusion proteins
Sustainable Development Goals
Citation
Rutkowska, DA, Meyer, QC, Maree, F, Vosloo, W, Fick, W & Huismans, H 2011, 'Use of soluble African horse sickness viral protein 7 as an antigen delivery and presentation system', Virus Research, vol. 156, no. 1-2, pp. 35-48.